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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 J+Am+Soc+Mass+Spectrom
2015 ; 26
(3
): 453-9
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Top-down analysis of highly post-translationally modified peptides by Fourier
transform ion cyclotron resonance mass spectrometry
#MMPMID25404158
Guerrero A
; Lerno L
; Barile D
; Lebrilla CB
J Am Soc Mass Spectrom
2015[Mar]; 26
(3
): 453-9
PMID25404158
show ga
Bovine ?-caseinoglycomacropeptide (GMP) is a highly modified peptide from
?-casein produced during the cheese making process. The chemical nature of GMP
makes analysis by traditional proteomic approaches difficult, as the peptide
bears a strong net negative charge and a variety of post-translational
modifications. In this work, we describe the use of electrospray ionization
Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR MS) for
the top-down analysis of GMP. The method allows the simultaneous detection of
different GMP forms that result from the combination of amino acid genetic
variations and post-translational modifications, specifically phosphorylation and
O-glycosylation. The different GMP forms were identified by high resolution mass
spectrometry in both negative and positive mode and confirmation was achieved by
tandem MS. The results showed the predominance of two genetic variants of GMP
that occur as either mono- or bi-phosphorylated species. Additionally, these four
forms can be modified with up to two O-glycans generally sialylated. The results
demonstrate the presence of glycosylated, bi-phosphorylated forms of GMP never
described before.